Impact of Starmerella bacillaris and Zygosaccharomyces bailii on ethanol reduction and Saccharomyces cerevisiae metabolism during mixed wine fermentations

The bulk of grape juice fermentation is carried out by the yeast Saccharomyces cerevisiae, but non-Saccharomyces yeasts can modulate many sensorial aspects of the final products in ways not well understood. In this study, some of such non-conventional yeasts were screened as mixed starter cultures in a fermentation defined medium in both simultaneous and sequential inoculations. One strain of Starmerella bacillaris and another of Zygosaccharomyces bailii were chosen by their distinct phenotypic footprint and their ability to reduce ethanol levels at the end of fermentation, particularly during simultaneous vinification. S. bacillaris losses viability strongly at the end of mixed fermentation, while Z. bailii remains viable until the end of vinification. Interestingly, for most non-Saccharomyces yeasts, simultaneous inoculation helps for survival at the end of fermentation compared to sequential inoculation. S. cerevisiae viability was unchanged by the presence of the either yeast. Characterization of both strains indicates that S. bacillaris behavior is overall more different from S. cerevisiae than Z. bailii. S. bacillaris has a less strict glucose repression mechanism and molecular markers like catabolite repression kinase Snf1 is quite different in size. Besides, S. cerevisiae transcriptome changes to a bigger degree in the presence of S. bacillaris than when inoculated with Z. bailii. S. bacillaris induces the translation machinery and repress vesicular transport. Both non-Saccharomyces yeast induce S. cerevisiae glycolytic genes, and that may be related to ethanol lowering, but there are specific aspects of carbon-related mechanisms between strains: Z. bailii presence increases the stress-related polysaccharides trehalose and glycogen while S. bacillaris induces gluconeogenesis genes.

Antimicrobial Activity of Nanoencapsulated Essential Oils of Tasmannia lanceolata, Backhousia citriodora and Syzygium anisatum against Weak-Acid Resistant Zygosaccharomyces bailii in Clear Apple Juice

The anti-yeast activity of oil-in-water encapsulated nanoemulsion containing individual or a combination of the three essential oils of Tasmanian pepper leaf (Tasmannia lanceolata), lemon myrtle (Backhousia citriodora), and anise myrtle (Syzygium anisatum) against weak-acid resistant Zygosaccharomyces bailii in clear apple juice was investigated. The effectiveness of the shelf-life extension of Z. bailii-spiked (1 × 103 CFU/mL) clear apple juice was evaluated and compared between natural (essential oils) and synthetic (sodium benzoate) antimicrobial agents. Essential oils showed an immediate reduction in the Z. bailii cell population at day-0 and exerted a fungicidal activity at day-4 of storage, with no further noticeable growth at the end of the experiment (day-28). At lower concentrations, Tasmanian pepper leaf oil of 0.0025% had >6 log CFU/mL at day-12 of storage. For lemon myrtle essential oils, the yeast population reached >6 log CFU/mL at day-24 and day-20 for concentrations of 0.02% and 0.01%, respectively. The fungicidal activity of Tasmanian pepper leaf oil reduced from 0.005% to 0.0025% v/v when mixed at a ratio of 1:1 with anise myrtle oil. The results of the present study suggest that these three native Australian herbs have the potential to be used in the beverage industry by controlling Zygosaccharomyces bailii in clear apple juice products.

Lemongrass and cinnamon oil nanoemulsions: Formulation and study of their physical stability and activity against Zygosaccharomyces bailii

The optimal conditions for elaborating oil/water nanoemulsions of lemongrass (LG), cinnamon bark (CB) and cinnamon leaves (CL) essential oils and their antimicrobial activity against Zygosaccharomyces bailii at pH 4.00 were studied. The effect of the emulsification methodology on the physical stability and antimicrobial activity of the nanoemulsions were also evaluated. Furthermore, the sensory impact of nanoemulsions added to an apple juice was tested. LG and CL nanoemulsions were elaborated by ultrasonication and CB nanoemulsion, by high-speed homogenization. They were stable for at least 120 days at 25 °C. They exhibited antimicrobial activity against Z. bailii being CB the most effective since it showed the smallest MIC value (156.3 mg/l), followed by LG (468.8 mg/l) and CL (1250.0 mg/l). A slight increase in growth rate was observed due to ultrasonication. An additive interaction in relation to the inhibitory effect between LG and CB nanoemulsions against Z. bailii was observed. While nanoemulsions obtained would be used as natural antimicrobial agents in food and beverage products, only LG nanoemulsion at MIC concentration diluted was acceptable in juice showing the sensory impact of essential oils on foods.

The N.C.Yeastract and CommunityYeastract databases to study gene and genomic transcription regulation in non-conventional yeasts

ABSTRACT Responding to the recent interest of the yeast research community in non-Saccharomyces cerevisiae species of biotechnological relevance, the N.C.Yeastract (http://yeastract-plus.org/ncyeastract/) was associated to YEASTRACT + (http://yeastract-plus.org/). The YEASTRACT + portal is a curated repository of known regulatory associations between transcription factors (TFs) and target genes in yeasts. N.C.Yeastract gathers all published regulatory associations and TF-binding sites for Komagataellaphaffii (formerly Pichia pastoris), the oleaginous yeast Yarrowia lipolytica, the lactose fermenting species Kluyveromyces lactis and Kluyveromyces marxianus, and the remarkably weak acid-tolerant food spoilage yeast Zygosaccharomyces bailii. The objective of this review paper is to advertise the update of the existing information since the release of N.C.Yeastract in 2019, and to raise awareness in the community about its potential to help the day-to-day work on these species, exploring all the information available in the global YEASTRACT + portal. Using simple and widely used examples, a guided exploitation is offered for several tools: (i) inference of orthologous genes; (ii) search for putative TF binding sites and (iii) inter-species comparison of transcription regulatory networks and prediction of TF-regulated networks based on documented regulatory associations available in YEASTRACT + for well-studied species. The usage potentialities of the new CommunityYeastract platform by the yeast community are also discussed.

Study of the Performance of Particles Based on Modified Starches Containing Potassium Sorbate and Incorporated into Biodegradable Films: Physicochemical Characterization and Antimicrobial Action

Ultrasound technique was used to produce native and acetylated cassava starch particles containing potassium sorbate (KS). In order to obtain an active packaging, films with addition of native starch particles containing KS (NKSPF) or added with acetylated starch particles containing KS (AKSPF) were formulated. As control systems, films without KS (CF) or added with KS that was not retained in particles (KSF), were produced. The NKSPF and AKSPF microstructure was consistent with composite materials. Tensile test revealed that CF and KSF were ductile and extensible (stress at break (σb) 2.8–2.5 MPa and strain at break (εb) 284–206%), while NKSPF and AKSPF were more resistant films with higher Young’s Modulus (148–477 MPa) and σb (3.6–17 MPa) but lower εb (40–11%). Moreover, NKSPF and AKSPF developed lower Yellowness Index (6.6–6.5) but higher opacity (19–23%) and solubility in water (31–35%) than KSF (9, 10.8% and 9%, respectively). It was observed that KSF and NKSPF moderately reduced the Zygosaccharomyces bailii growth while AKSPF showed the highest yeast inhibition, three Log-cycles, compared to CF. Additionally, FTIR spectroscopy revealed intensified interactions between KS and modified starch. It was concluded that starch sonication and acetylation were useful modifications to produce particles carrier of KS that improved the physical and antimicrobial performance of active films.

The Use of Yeast Mixed Cultures for Deacidification and Improvement of the Composition of Cold Climate Grape Wines

Interest in the use of non-Saccharomyces yeast in mixed cultures is increasing due to the perceived improvement in the quality and complexity of the resulting wines. The aim of the study was to determine the ability of monocultures and mixed yeast cultures for deacidification and improvement of the composition of cold climate grape wines. Fermentation of grape musts with increased total acidity was carried out with the use of monocultures of Saccharomyces cerevisiae MH020215 (Sc), Zygosaccharomyces bailii 749 (Zb) and Metschnikowia pulcherrima MG970690 (Mp), and their mixed cultures, inoculated simultaneously and sequentially. Oenological parameters, organic acids and volatile compounds profiles of obtained wines were characterized. The fermentation kinetics and analytical profiles of the obtained wines showed that the use of mixed yeast cultures contributed to the reduction of volatile acidity and acetic acid content in the wines, as well as obtaining a favorable aromatic profile of the wines. The dominant higher alcohols in all wines were 2-methyl-1-propanol, 3-methyl-1-butanol and 2-methyl-1-butanol. Significantly higher amounts of the first two compounds were found in wines obtained with M. pulcherrima MG070690, both in monoculture and in mixed cultures. The monocultures of M. pulcherrima MG070690 (Mp) compared with Z. bailli 749 (Zb) synthesized higher levels of esters in wines, including ethyl acetate, ethyl propionate, isobutyl acetate, ethyl pyroracemate and isoamyl acetate.

Effect of a Multistarter Yeast Inoculum on Ethanol Reduction and Population Dynamics in Wine Fermentation

Microbiological strategies are currently being considered as methods for reducing the ethanol content of wine. Fermentations started with a multistarter of three non-Saccharomyces yeasts (Metschnikowia pulcherrima (Mp), Torulaspora delbrueckii (Td) and Zygosaccharomyces bailii (Zb)) at different inoculum concentrations. S. cerevisiae (Sc) was inoculated into fermentations at 0 h (coinoculation), 48 h or 72 h (sequential fermentations). The microbial populations were analyzed by a culture-dependent approach (Wallerstein Laboratory Nutrient (WLN) culture medium) and a culture-independent method (PMA-qPCR). The results showed that among these three non-Saccharomyces yeasts, Td became the dominant non-Saccharomyces yeast in all fermentations, and Mp was the minority yeast. Sc was able to grow in all fermentations where it was involved, being the dominant yeast at the end of fermentation. We obtained a significant ethanol reduction of 0.48 to 0.77% (v/v) in sequential fermentations, with increased concentrations of lactic and acetic acids. The highest reduction was achieved when the inoculum concentration of non-Saccharomyces yeast was 10 times higher (107 cells/mL) than that of S. cerevisiae. However, this reduction was lower than that obtained when these strains were used as single non-Saccharomyces species in the starter, indicating that interactions between them affected their performance. Therefore, more combinations of yeast species should be tested to achieve greater ethanol reductions.

Antioxidative, Antifungal and Additive Activity of the Antimicrobial Peptides Leg1 and Leg2 from Chickpea

The fight against food waste benefits from novel agents inhibiting spoilage. The present study investigated the preservative potential of the antimicrobial peptides Leg1 (RIKTVTSFDLPALRFLKL) and Leg2 (RIKTVTSFDLPALRWLKL) recently identified in chickpea legumin hydrolysates. Checkerboard assays revealed strong additive antimicrobial effects of Leg1/Leg2 with sodium benzoate against Escherichia coli and Bacillus subtilis with fractional inhibitory concentrations of 0.625 and 0.75. Additionally, Leg1/Leg2 displayed antifungal activity with minimum inhibitory concentrations of 500/250 µM against Saccharomyces cerevisiae and 250/125 µM against Zygosaccharomyces bailii. In contrast, no cytotoxic effects were observed against human Caco-2 cells at concentrations below 2000 µM (Leg1) and 1000 µM (Leg2). Particularly Leg2 showed antioxidative activity by radical scavenging and reducing mechanisms (maximally 91.5/86.3% compared to 91.2/94.7% for the control ascorbic acid). The present results demonstrate that Leg1/Leg2 have the potential to be applied as preservatives protecting food and other products against bacterial, fungal and oxidative spoilage.

Performance of Wild Non-Conventional Yeasts in Fermentation of Wort Based on Different Malt Extracts to Select Novel Starters for Low-Alcohol Beers

Nowadays, the increasing interest in new market demand for alcoholic beverages has stimulated the research on useful strategies to reduce the ethanol content in beer. In this context, the use of non-Saccharomyces yeasts to produce low-alcohol or alcohol-free beer may provide an innovative approach for the beer market. In our study, four wild non-Saccharomyces yeasts, belonging to Torulaspora delbrueckii, Candida zemplinina and Zygosaccharomyces bailii species, were tested in mixed fermentation with a wild selected Saccharomyces cerevisiae strain as starters for fermentation of different commercial substrates used for production of different beer styles (Pilsner, Weizen and Amber) to evaluate the influence of the fermentative medium on starter behaviour. The results obtained showed the influence of non-Saccharomyces strains on the ethanol content and organoleptic quality of the final beers and a significant wort–starter interaction. In particular, each starter showed a different sugar utilization rate in each substrate, in consequence of uptake efficiency correlated to the strain-specific metabolic pathway and substrate composition. The most suitable mixed starter was P4-CZ3 (S. cerevisiae–C. zemplinina), which is a promising starter for the production of low-alcohol beers with pleasant organoleptic characteristics in all the tested fermentation media.

Study of the effect of zygosaccharomyces bailii on microbiological spoilage of food products

The article presents the results of studies of the influence of breeding conditions and properties of Zygosaccharomyces bailii on microbiological spoilage of food products. Particular attention is paid to the resistance of this microorganism to various conditions and its resistance to sanitization.